Protamine Sulfate Enhances Biofilm Inhibitory Activity of Curcumin against Pseudomonas Aeruginosa Biofilm Grown on Ex Vivo Porcine Skin Model

Authors

  • Solaiman Edeas Department of Applied Pharmaceutical sciences, Faculty of pharmacy, Isra University, Amman, Jordan
  • Nisreen Ahmad Dahshan Department of Applied Pharmaceutical sciences, Faculty of pharmacy, Isra University, Amman, Jordan
  • Ahmad Talhouni Department of Applied Pharmaceutical sciences, Faculty of pharmacy, Isra University, Amman, Jordan
  • Suha Mujahed Abudoleh Department of Basic Pharmaceutical sciences, Faculty of pharmacy, Isra University, Amman, Jordan, Head of the Department of Basic Pharmaceutical Sciences, Faculty of Pharmacy- Isra University, Amman- Jordan

Keywords:

Biofilm, Curcumin, Protamine sulfate, Quorum sensing inhibitors, Pseudomonas aeruginosa

Abstract

A bacterial biofilm is one of the most difficult structures to eradicate and is involved in the enhancement of the virulence and resistance of bacteria. Several strategies have been used to fight bacterial virulence and biofilm formation through targeting the quorum sensing (QS) mechanism of communication. Curcumin is a natural component extract from Turmeric and has antimicrobial and quorum sensing inhibitory (QSI) effects on many microbes and their biofilm. This study aimed to evaluate the effect of curcumin-protamine sulfate combination against the biofilm formation by P. aeruginosa (ATCC 27853). Neither the curcumin nor the protamine sulfate exhibited antibiofilm activity when tested against the P .aeruginosa biofilm alone. However, when curcumin was combined with protamine sulfate at different concentrations, a significant reduction in biofilm formation was detected. The highest inhibition percentage was detected against 48 hour biofilm when the biofilm was treated with 62.5 µg/ml curcumin with 62.5 µg/ml protamine sulfate using the 96-well plate method. The porcine ex-vivo model was used to confirm the previous result. The highest inhibition of biofilm was 95% when the porcine skin was treated with 500 µg/ml of curcumin with 500 µg/ml protamine sulfate. The obtained results from this study highly suggest that the combination of curcumin with protamine sulfate improves the effect against the biofilm, and ensures that the effect is due to the prevention of biofilm formation mainly through interruption of QS rather than killing of the bacteria.

References

Bjarnsholt, T. (2013) The role of bacterial biofilms in chronic infections. APMIS. Supplementum. (136), 1–51.

Xu, Z. and Hsia, H.C. (2018) The impact of microbial communities on wound healing: A review. Annals of Plastic Surgery. 81 (1), 113–123.

Brandenburg, K.S., Weaver, A.J., Karna, S.L.R., You, T., Chen, P., Stryk, S. Van, et al. (2019) Formation of Pseudomonas aeruginosa Biofilms in Full-thickness Scald Burn Wounds in Rats. Scientific Reports. 9 (1), 1–12.

Serra, R., Grande, R., Butrico, L., Rossi, A., Settimio, U.F., Caroleo, B., et al. (2015) Chronic wound infections: The role of Pseudomonas aeruginosa and Staphylococcus aureus. Expert Review of Anti-Infective Therapy. 13 (5), 605–613.

Lee, K. and Yoon, S.S. (2017) Pseudomonas aeruginosa Biofilm, a Programmed Bacterial Life for Fitness. Journal of Microbiology and Biotechnology. 27 (6), 1053–1064.

Thi, M.T.T., Wibowo, D., and Rehm, B.H.A. (2020) Pseudomonas aeruginosa biofilms. International Journal of Molecular Sciences. 21 (22), 1–25.

Wei, Q. and Ma, L.Z. (2013) Biofilm matrix and its regulation in Pseudomonas aeruginosa. International Journal of Molecular Sciences. 14 (10), 20983–21005.

Mulcahy, L.R., Isabella, V.M., and Lewis, K. (2013) Pseudomonas aeruginosa Biofilms in Disease.

Wang, H., Chu, W., Ye, C., Gaeta, B., Tao, H., Wang, M., et al. (2019) Chlorogenic acid attenuates virulence factors and pathogenicity of Pseudomonas aeruginosa by regulating quorum sensing. Applied Microbiology and Biotechnology. 103 (2), 903–915.

Mielko, K.A., Jab?o?ski, S.J., Milczewska, J., Sands, D., ?ukaszewicz, M., and M?ynarz, P. (2019) Metabolomic studies of Pseudomonas aeruginosa. World Journal of Microbiology and Biotechnology. 35 (11), 1–11.

Pang, Z., Raudonis, R., Glick, B.R., Lin, T.J., and Cheng, Z. (2019) Antibiotic resistance in Pseudomonas aeruginosa: mechanisms and alternative therapeutic strategies. Biotechnology Advances. 37 (1), 177–192.

Moradali, M.F., Ghods, S., and Rehm, B.H.A. (2017) Pseudomonas aeruginosa lifestyle: A paradigm for adaptation, survival, and persistence. Frontiers in Cellular and Infection Microbiology. 7 (FEB),.

Nelson, K.M., Dahlin, J.L., Bisson, J., Graham, J., Pauli, G.F., and Walters, M.A. (2017) The Essential Medicinal Chemistry of Curcumin. Journal of Medicinal Chemistry. 60 (5), 1620–1637.

Pulido-Moran, M., Moreno-Fernandez, J., Ramirez-Tortosa, C., and Ramirez-Tortosa, M.C. (2016) Curcumin and health. Molecules. 21 (3), 1–22.

Rai, M., Ingle, A.P., Pandit, R., Paralikar, P., Anasane, N., and Santos, C.A. Dos (2020) Curcumin and curcumin-loaded nanoparticles: antipathogenic and antiparasitic activities. Expert Review of Anti-Infective Therapy. 18 (4), 367–379.

Bhawana, Basniwal, R.K., Buttar, H.S., Jain, V.K., and Jain, N. (2011) Curcumin nanoparticles: Preparation, characterization, and antimicrobial study. Journal of Agricultural and Food Chemistry. 59 (5), 2056–2061.

Praditya, D., Kirchhoff, L., Brüning, J., Rachmawati, H., Steinmann, J., and Steinmann, E. (2019) Anti-infective properties of the golden spice curcumin. Frontiers in Microbiology. 10 (MAY), 1–16.

Neyestani, Z., Ebrahimi, S.A., Ghazaghi, A., Jalili, A., Sahebkar, A., and Rahimi, H.R. (2019) Review of anti-bacterial activities of curcumin against pseudomonas aeruginosa. Critical Reviews in Eukaryotic Gene Expression. 29 (5), 377–385.

Jahromi, M.A.M., Al-Musawi, S., Pirestani, M., Ramandi, M.F., Ahmadi, K., Rajayi, H., et al. (2014) Curcumin-loaded chitosan tripolyphosphate nanoparticles as a safe, natural and effective antibiotic inhibits the infection of staphylococcus aureus and pseudomonas aeruginosa in vivo. Iranian Journal of Biotechnology. 12 (3),.

Mukhopadhyay, K., Kumari, A., Tyagi, P., Singh, M., and Kumari, H. (2015) Bactericidal Activity of Curcumin I Is Associated with Damaging of Bacterial Membrane. Plos One. 10 (3), e0121313.

Niamsa, N. and Sittiwet, C. (2009) Antimicrobial activity of Curcuma longa aqueous extract. Journal of Pharmacology and Toxicology. 4 (4), 173–177.

Gholami, M., Zeighami, H., Bikas, R., Heidari, A., Rafiee, F., and Haghi, F. (2020) Inhibitory activity of metal-curcumin complexes on quorum sensing related virulence factors of Pseudomonas aeruginosa PAO1. AMB Express. 10 (1),.

Xue, B., Huang, J., Zhang, H., Li, B., Xu, M., Zhang, Y., et al. (2020) Micronized curcumin fabricated by supercritical CO2 to improve antibacterial activity against Pseudomonas aeruginosa. Artificial Cells, Nanomedicine and Biotechnology. 48 (1), 1135–1143.

Prateeksha, Rao, C. V., Das, A.K., Barik, S.K., and Singh, B.N. (2019) ZnO/Curcumin Nanocomposites for Enhanced Inhibition of Pseudomonas aeruginosa Virulence via LasR-RhlR Quorum Sensing Systems. Molecular Pharmaceutics. 16 (8), 3399–3413.

Rahbar Takrami, S., Ranji, N., and Sadeghizadeh, M. (2019) Antibacterial effects of curcumin encapsulated in nanoparticles on clinical isolates of Pseudomonas aeruginosa through downregulation of efflux pumps. Molecular Biology Reports. 46 (2), 2395–2404.

Soleimani, V. and Sahebkr, Amirhossein Hosseinzadeh, H. (2018) Turmeric (Curcuma longa) and its major constituent (curcumin) as nontoxic and safe substances: Review. Phytotherapy Research. 32 (6), 985–995.

Aziz, M., Garduno, R., Mirani, Z.A., Baqai, R., Sheikh, A.S., Nazir, H., et al. (2019) Determination of antimicrobial effect of protamine by transmission electron microscopy and SDS PAGE on Pseudomonas aeruginosa isolates from diabetic foot infection. Iranian Journal of Basic Medical Sciences. 22 (7), 827–832.

Soboh, F., Khoury, A.E., Zamboni, A.C., Davidson, D., and Mittelman, M.W. (1995) Effects of ciprofloxacin and protamine sulfate combinations against catheter-associated Pseudomonas aeruginosa biofilms. Antimicrobial Agents and Chemotherapy. 39 (6), 1281–1286.

Yakandawala, N., Gawande, P. V., LoVetri, K., and Madhyastha, S. (2007) Effect of ovotransferrin, protamine sulfate and EDTA combination on biofilm formation by catheter-associated bacteria. Journal of Applied Microbiology. 102 (3), 722–727.

Boussard, P., Devleeschouwer, M., and Dony, J. (1994) Influence of protamine on the in vitro sensitivity of Pseudomonas aeruginosa to antibiotics. Pharmaceutica Acta Helvetiae. 68 (3), 161–167.

Darouiche, R.O., Mansouri, M.D., Gawande, P. V., and Madhyastha, S. (2008) Efficacy of combination of chlorhexidine and protamine sulphate against device-associated pathogens. Journal of Antimicrobial Chemotherapy. 61 (3), 651–657.

Dahshan, N.A., Abudoleh, S.M., Talhouni, A., and Alkhani, Z. (2021) Evaluation of Pseudomonas aeruginosa Antibiofilm Activity of Chlorogenic Acid - Protamine Sulfate Combination Using Ex Vivo Porcine Skin Model. Journal of Hunan University Natural Sciences. 48 (12),.

Wedajo, W., Schön, T., Bedru, A., Kiros, T., Hailu, E., Mebrahtu, T., et al. (2014) A 24-well plate assay for simultaneous testing of first and second line drugs against Mycobacterium tuberculosis in a high endemic setting. BMC Research Notes. 7 (1), 1–8.

Chen, H., Wubbolts, R.W., Haagsman, H.P., and Veldhuizen, E.J.A. (2018) Inhibition and Eradication of Pseudomonas aeruginosa Biofilms by Host Defence Peptides. (March), 1–10.

Phillips, P.L., Yang, Q., Davis, S., Sampson, E.M., Azeke, J.I., Hamad, A., et al. (2015) Antimicrobial dressing efficacy against mature Pseudomonas aeruginosa biofilm on porcine skin explants. International Wound Journal. 12 (4), 469–483.

Yang, Q., Phillips, P.L., Sampson, E.M., Progulske-Fox, A., Jin, S., Antonelli, P., et al. (2013) Development of a novel ex vivo porcine skin explant model for the assessment of mature bacterial biofilms. Wound Repair and Regeneration. 21 (5), 704–714.

Rasamiravaka, T., Labtani, Q., Duez, P., and El Jaziri, M. (2015) The formation of biofilms by pseudomonas aeruginosa: A review of the natural and synthetic compounds interfering with control mechanisms. BioMed Research International. 2015.

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Published

2022-10-28

How to Cite

Solaiman Edeas, Nisreen Ahmad Dahshan, Talhouni, A., & Suha Mujahed Abudoleh. (2022). Protamine Sulfate Enhances Biofilm Inhibitory Activity of Curcumin against Pseudomonas Aeruginosa Biofilm Grown on Ex Vivo Porcine Skin Model. International Journal of Natural Sciences: Current and Future Research Trends, 16(1), 31–40. Retrieved from https://ijnscfrtjournal.isrra.org/index.php/Natural_Sciences_Journal/article/view/1133

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